Fig. 7: Enhanced NRP1/KDR/EFNB2 expression as a marker set of venous arterialization of the VECs in PVOD patient lung.
From: Macrophage ferroptosis potentiates GCN2 deficiency induced pulmonary venous arterialization
a Representative images of NRP1/KDR/EFNB2 (red), NR2F2 (yellow), CD31 (green), and α-SMA (cyan) immunofluorescence and DAPI (blue) staining of lung tissues from Control and PVOD patients, with the boxed region magnified. Scale bar = 50 µm. Enlarged view scale bar = 10 µm. The data are representative images. Control, n = 7 individuals; PVOD n = 9 individuals with similar results. b qRT-PCR analyzed the expression levels of representative markers at various stages of VECs differentiation process. Data are presented as the means ± s.e.m. (n = 4 replicates); two-way ANOVA with Tukey’s multiple comparison test. CD31 VEC, p = 6 × 10−12; NR2F2 VEC, p = 4 × 10−10. c qRT-PCR analyzed the differential expression levels of candidate genes in Control-iPSC VEC (Con) and PVOD-iPSC VEC (PVOD). Data are the mean ± s.e.m. (n = 6), unpaired two-sided t-test. EFNB2 Con vs PVOD, p = 4.7 × 10−4; KDR Con vs PVOD, p = 8 × 10−6. d Immunofluorescence analysis of a venous marker (NR2F2) with DAPI counterstaining. Scale bar = 100 µm. The experiment was repeated independently four times with similar results. e Western blot showing the expression levels of NRP1, EFNB2, p-ERK, Total-ERK in Control-iPSC VEC (Con) or PVOD iPSC-VEC (PVOD). The experiment was repeated independently three times with similar results. f Immunofluorescence analysis of p-ERK with DAPI counterstaining in Control-iPSC VEC (Con) or PVOD iPSC-VEC (PVOD). Scale bar = 50 µm. Data are the mean ± s.e.m. (n = 6 replicates), unpaired two-sided t-test. Each dot represents an individual biological replicate, at least three independent experiments. P values are indicated in the figures. Source data are provided as a Source data file.
